Evaluating the reliability of microsatellite genotyping from low-quality DNA templates with a polynomial distribution model

• 来源期刊: 科学通报(英文版)
• 学科: 生物学
• 年，卷（期）: 2011,（24）
• 页码范围: 2523-2530
• 页数: 8页
• 分类号: Q75
• 语种: 中文

摘要

Molecular studies using trace DNA, such as from museum specimens, ancient or forensic samples and samples obtained noninvasively, often have a common problem of low quality of DNA templates. Amplification errors, such as allelic dropout and false allele, may arise during polymerase chain reaction (PCR) using such samples. A mathematical model which treats homozygotes and heterozygotes discriminately has been developed to measure sample quality and compute the confidence level of using multipletube approaches. We use plucked hair samples collected from 26 individual Sichuan snub-nosed monkeys (Rhinopithecus roxel- lana) to test the model. In this case, a confidence level of 99% can be achieved by three positive PCRs. If the sample quality is very poor and requires many PCR replicates, an alternative multiplestep genotyping method is recommended. This model enables researchers to optimize experimental protocols through pilot studies and obtain reliable genetic information using noninvasive sampling method.